施氏假单胞菌JHY01菌株毒死蜱降解酶的定位及其提取条件的优化  

Localization and Extraction Conditions of Chlorpyrifos-Degrading Enzyme from Pseudomonas stutzeri

作　　者：兰亚红[1] 谢明[1] 陈福良[1] 万方浩[1] 周淑云[1] 

机构地区：[1]中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室,北京100094

出　　处：《中国生物防治》2008年第4期349-353,共5页Chinese Journal of Biological Control

基　　金：国家科技支撑计划(2007BDA87B08;2006BAD08A02;2006BAD08A08);国家863计划(2006AA10Z401)

摘　　要：对具有降解有机磷农药毒死蜱作用的施氏假单胞菌JHY01菌株的降解酶进行定位,测得其胞外酶、胞内酶、细胞碎片中残留酶对毒死蜱的降解率分别为8.41%、79.85%、77.14%。由此确定该菌株的有机磷降解酶主要为胞内酶。采用超声波破碎方法提取降解酶,对降解酶提取条件进行正交优化,结果显示,最佳超声波(超声波碎仪Sonics-VCX500,功率500W,频率20kHz)破碎条件是:菌体重量:缓冲液体积为1:4(g/ml),总辐射次数30次,每次辐射4s,间隔4s。在此条件下所提取的粗酶液酶活力最高,对毒死蜱的降解效率可达84.47%。Pseudomonas stutzeri JHY01 was capable of degrading Chlorpyrifos, a typical organophosphorus pesticide. The ultrasonic cell-break method was used to extract the degrading enzyme. Results showed the degradation rates of chlorpyrifos by extracellular enzyme, intracellular enzyme and cell fragment were 8.41%, 79.85% and 77.14%, respectively, indicating that Chlorpyrifos-degrading enzymes were typi- cal intracellular enzymes. Orthogonal experiments were then designed to optimize the extraction conditions of the degrading enzymes. The degrading activity of the crude enzymes was maximal （with the degradation rate 84.47 % ） when the ultrasonic conditions were as follows ： the biomass of the cells to extraction buffer volume 1 ： 4 （g/ml）, with ultrasonic treatment for 4 seconds, totally 30 times at interval of 4 seconds （UAS, Sonics-VCX500, 500W, 25KHz）.

关 键 词：施氏假单胞菌 降解酶 酶定位 提取条件优化 超声波破碎法 

分 类 号：X592[环境科学与工程—环境工程] X172